This SBIR Phase I proposal aims to develop and commercialize a portable, easy-to-use system for nucleic acid preparation for effective molecular diagnostics in low-resource settings (LRS). The technology will also aid in the monitoring of treatment response for patients of global diseases such as AIDS, malaria, and tuberculosis. Because a reliable and reproducible high-quality sample preparation process for NA is critical to the success of most molecular diagnostic assays and the subsequent treatment and monitoring of recovery progress, the availability of an effective sample preparation technology is especially important if the tests are performed in non-traditional or low-resource health care settings. Most processes in current commercial sample preparation methods are labor intensive and require special instrumentation. Although commercially available automated systems can perform extensive sample processing, they are expensive for use in non-central laboratory settings. Several research efforts have focused on the development of a sample in, answer out microfluidic total analysis system (TAS). While these efforts have often resulted in TAS platforms with impressive analytical capabilities and represent a potentially paradigm-shifting long term solution, they remain an area of active research and may not be applicable to LRS settings. For instance, the use of these systems is inhibited by their continuing need for instrumentation that is too sophisticated and costly for LRS. To this end, a portable, centrifuge-free, room temperature nucleic acid (NA) extraction system has recently been reported by our collaborator. This system for nucleic acid preparation (or SNAP) requires only thermally stable reagents so the system is compatible with storage and transport in LRS. Unlike other sample preparation technology in which the captured NA needs to be eluted immediately, samples are stabilized on detachable cartridges, which can be eluted either at the point of care, or transported to a centralized laboratory for later analysis. Using a patent pending, multi-chamber, and prefilled syringe technology, AI Biosciences, Inc. (AIBI) proposes to significantly reduce the size and weight of the SNAP technology and to reduce the total hands-on time needed to perform an extraction. This single, pre-filled syringe will contain all the reagents required to conduct DNA/RNA extraction via the solid phase extraction (SPE) column in the SNAP. The actuation of a single plunger can sequentially deliver the necessary reagents to this SPE column to perform an extraction. In Phase I, we will demonstrate effective extraction from realistic samples and to deliver PCR and RT-PCR ready templates in 15 min or less. The yield and purity of the isolated NA are expected to match those obtained from commercially available kits that require far more instruments and hands-on time. We will also assess the long term stability of the pre-filled reagents for practical use in LRS. If the work in Phase I is successful we will seek to develop a low-cost, fully automated, and multi-sample system in Phase II.